Multiplex Tissue Analysis

Analyzing multiple biomarkers in tissues with preserved anatomical context presents compelling opportunities to uncover crucial insights into the underlying mechanisms driving diverse disease processes. This approach is increasingly gaining traction in clinical research and among pharmaceutical teams, serving as invaluable tools for target validation, efficacy analyses, and patient stratification.

However, executing such analyses is a complex undertaking, demanding significant investments in expensive infrastructure, finely calibrated and integrated workflows, and highly trained staff. This challenge becomes even more pronounced when handling larger cohorts of tissues that require meticulous processing and quantitative analysis.

At Offspring, we offer a unique capability and capacity to efficiently co-process large quantities of tissues for the analysis of multiple biomarkers. Leveraging multiplexed immunofluorescence or immunohistochemical , as well as Nanostring-based analyses, along with pre-established or custom-built biomarker panels, we combine these capabilities with agile digital scanning of stained tissue sections. Our approach incorporates deep learning AI-powered quantitative image analysis, ensuring the generation of reliable data crucial for making informed stop/go decisions. This comprehensive solution is designed to meet the rigorous demands of spatial biology research, providing a streamlined and effective means to extract valuable insights from tissue analyses."

Multiplex Tissue Analysis: About Us

Offspring's Preestablished Panels

Offspring Biosciences is set up to assist your projects with co-staining of multiple biomarkers in tissue sections using a range of techniques, including up to 6-plexed immunohistofluorescence (IHF), 3-plexed chromogenic immunohistochemistry (IHC) and 2-plexed in situ hybridization histochemistry (ISH). We perform these analyses with pre-established panels for multiplexed biomarker staining or by custom build to flexibly adapt their design to your specific needs.

Listed below are examples of the multiplexed analyses we can run for you:

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CD8 and NK Cell Activity

CD8
Ki67
NKp46
Granzyme B

This 4-plex-antibody panel enables us to evaluate if your drug triggers cytotoxic T and NK cells infiltration. This can be used in preclinical treatment studies of syngeneic mouse models for cancer. More specifically, the CD8 and NKp46 are markers for cytotoxic T cells and NK cells, respectively. Granzyme B is one of the T and NK cytotoxic cell effector enzymes used to kill cells (eg tumor cells). Ki67 identifies cells in proliferation; for instance, showing if the treatment influences T or NK cells’ proliferation.

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CD4 Cell Activity

CD4
PD-1
Tim-3
FoxP3

The CD4 cell activity antibody panel is used to identify and profile the functional state of CD4-positive T cells. For instance, this can be used in the context of mouse syngeneic tumor environments. More specifically, cells marked with only CD4 are identified as T helper cells. When CD4 cells also express FoxP3 they are acting in an immunosuppressive T regulatory state. PD-1 and TIM-3 are both cell surface receptors actively induced on T cells after antigenic stimuli. They can indicate the immunosuppressed/exhausted phenotype of either CD4 or CD8 cells.

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Tumor Phenotype and Activity

Ki67
Vimentin
Epithelial
Activity Marker

This 4-plex antibody panel allows us to state the phenotype and activity of a mouse or human tumor as means to evaluate the effect of a drug in treatment studies. In this panel, antibodies identify the tumor’s epithelial or mesenchymal-like phenotype. In pan-cytokeratin or vimentin positive tumor cells, the nuclear Ki-67 staining “transcriptional activity marker” show their activation and proliferation. Hoescht 33342 nuclear staining also provides information about the viability of the tumor cells. One of two serial sections can be stained with this panel and the results combined with an adjacent immune marker panel revealing the physical/anatomical relation between the tumor cell mass and invading immune cells.

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